Nongroupable Strains Research Articles

Meningococcal Carriage in Children with Atypical Hemolytic Uremic Syndrome Receiving Eculizumab Therapy.

Eculizumab is a first-line treatment for atypical hemolytic uremic syndrome (aHUS), and patients undergoing eculizumab therapy may become more susceptible to infection caused by Neisseria meningitidis (Nm). While meningococcal vaccination is required for patients undergoing eculizumab therapy, there is limited knowledge about meningococcal carriage in children with aHUS. We aimed to evaluate (1) the prevalence of Nm carriage, (2) serogroup distribution, and (3) the immunization status of children undergoing eculizumab treatment for aHUS. The Meningo-aHUS study is a prospective, multi-center study evaluating meningococcal carriage in children and adolescents in Türkiye receiving eculizumab for aHUS. We noted the age, gender, daycare, school, or university attendance, passive smoking status, previous infection and antibiotic use, and previous immunization history, including meningococcal vaccines, from the medical records of those children with aHUS. We collected nasopharyngeal samples, tested them for Nm using real-time polymerase chain reaction, and performed a serogroup analysis on the positive samples. We collected nasopharyngeal samples from 62 children with aHUS. Out of 62 children, 61 (98.4%) had received at least one dose of the meningococcal vaccine. The median time since the last meningococcal vaccine dose was 15 months (1-59 months). We detected meningococcal carriage in three (4.8%, 95% CI 1.0-13.5) children, and all three strains were non-groupable (NG). No other serogroups were detected. Almost all the children received their risk-group meningococcal immunization, including booster doses. A 4.8% of children with aHUS carried NG meningococci and, no vaccine serogroups were detected. Patients treated with eculizumab remain profoundly susceptible to IMD due to these NG meningococcal strains. The occurrence of breakthrough cases and carriage of Nm, especially NG strains, highlights the significance of maintaining a state of constant alertness, promptly seeking medical attention, and swiftly treating any symptoms that align with IMD, regardless of their vaccination status or antibiotic prophylaxis.

Invasive meningococcal disease in patients with complement deficiencies: a case series (2008\u20132017)

BackgroundTo describe patients with inherited and acquired complement deficiency who developed invasive meningococcal disease (IMD) in England over the last decade.MethodsPublic Health England conducts enhanced surveillance of IMD in England. We retrospectively identified patients with complement deficiency who developed IMD in England during 2008–2017 and retrieved information on their clinical presentation, vaccination status, medication history, recurrence of infection and outcomes, as well as characteristics of the infecting meningococcal strain.ResultsA total of 16 patients with 20 IMD episodes were identified, including four with two episodes. Six patients had inherited complement deficiencies, two had immune-mediated conditions associated with complement deficiency (glomerulonephritis and vasculitis), and eight others were on Eculizumab therapy, five for paroxysmal nocturnal haemoglobinuria and three for atypical haemolytic uraemic syndrome. Cultures were available for 7 of 11 episodes among those with inherited complement deficiencies/immune-mediated conditions and the predominant capsular group was Y (7/11), followed by B (3/11) and non-groupable (1/11) strains. Among patients receiving Eculizumab therapy, 3 of the 9 episodes were due to group B (3/9), three others were NG but genotypically group B, and one case each of groups E, W and Y.ConclusionsIn England, complement deficiency is rare among IMD cases and includes inherited disorders of the late complement pathway, immune-mediated disorders associated with low complement levels and patients on Eculizumab therapy. IMD due to capsular group Y predominates in patient with inherited complement deficiency, whilst those on Eculizumab therapy develop IMD due to more diverse capsular groups including non-encapsulated strains.

Characterization of strains of Neisseria meningitidis causing meningococcal meningitis in Mozambique, 2014: Implications for vaccination against meningococcal meningitis.

IntroductionIn sub Saharan Africa, the epidemiology, including the distribution of serogroups of strains of N. meningitidis is poorly investigated in countries outside “the meningitis belt”. This study was conducted with the aim to determine the distribution of serogroups of strains of N. meningitidis causing meningococcal meningitis in children and adults in Mozambique.MethodsA total of 106 PCR confirmed Neisseria meningitidis Cerebrospinal Fluid (CSF) samples or isolates were obtained from the biobank of acute bacterial meningitis (ABM) surveillance being implemented by the National Institute of Health, at three central hospitals in Mozambique, from January to December 2014. Serogroups of N. meningitidis were determined using conventional PCR, targeting siaD gene for Neisseria meningitidis. Outer Membrane Proteins (OMP) Genotyping was performed by amplifying porA gene in nine samples.ResultsOf the 106 PCR confirmed Neisseria meningitidis samples, the most frequent serotype was A (50.0%, 53/106), followed by W/Y (18.9%, 20/106), C (8.5%, 9/106), X (7.5%, 8/106) and B (0.9%, 1/106). We found non-groupable strains in a total of 15 (14.2%) samples. PorA genotypes from nine strains showed expected patterns with the exception of two serogroup C strains with P1.19,15,36 and P1.19–36,15 and one serogroup X with P1.19,15,36, variants frequently associated to serogroup B.ConclusionOur data shows that the number of cases of meningococcal meningitis routinely reported in central hospitals in Mozambique is significant and the most dominant serogroup is A. In conclusion, although serogroup A has almost been eliminated from the “meningitis belt”, this serogroup remains a major concern in countries outside the belt such as Mozambique.

Meningococcal carriage among a university student population – United States, 2015

ObjectivesSeveral outbreaks of serogroup B meningococcal disease have occurred among university students in recent years. In the setting of high coverage of the quadrivalent meningococcal conjugate vaccine and prior to widespread use of serogroup B meningococcal vaccines among adolescents, we conducted surveys to characterize the prevalence and molecular characteristics of meningococcal carriage among university students. MethodsTwo cross-sectional oropharyngeal carriage surveys were conducted among undergraduates at a Rhode Island university. Isolates were characterized using slide agglutination, real-time polymerase chain reaction (rt-PCR), and whole genome sequencing. Adjusted prevalence ratios and 95% confidence intervals were calculated using Poisson regression to determine risk factors for carriage. ResultsA total of 1837 oropharyngeal specimens were obtained from 1478 unique participants. Overall carriage prevalence was 12.7–14.6% during the two survey rounds, with 1.8–2.6% for capsular genotype B, 0.9–1.0% for capsular genotypes C, W, or Y, and 9.9–10.8% for nongroupable strains by rt-PCR. Meningococcal carriage was associated with being male, smoking, party or club attendance, recent antibiotic use (inverse correlation), and recent respiratory infections. ConclusionsIn this university setting, the majority of meningococcal carriage was due to nongroupable strains, followed by serogroup B. Further evaluation is needed to understand the dynamics of serogroup B carriage and disease among university students.

Prevalence of factor H Binding Protein sub-variants among Neisseria meningitidis in China

ObjectiveTo study the prevalence of the fHbp genes in Neisseria meningitidis (N. meningitidis) isolates for further evaluation and development of serogroup B meningococcal vaccines in China. MethodsA panel of 1012 N. meningitidis strains was selected from the national culture collection from 1956 to 2016, according to the years of isolation, locations, and strain sources. These were tested by FHbp variant typing. Multi-locus sequence typing (MLST) was performed on 822 of these samples, including 242 strains from clinical strains and 580 carrier-derived strains. Analysis based on sequence types, serogroups, and FHbp variations were used to summarize the prevalence and characteristics of N. meningitidis. ResultsThere were 8 serogroups of N. meningitidis as well as a collection of nongroupable strains in this study. 1008 of 1012 N. meningitidis strains tested were positive for the fHbp gene. Serogroup A N. meningitidis (MenA) strains belonging to ST-1 and ST-5 clonal complexes harbored genes only encoding variant 1 (v1) FHbp. All MenW strains encoded v2 FHbp. 61.9% of clinical MenB strains were positive for v2 FHbp vs. 32.1% that were positive for v1. Among fHbp-positive carrier-derived MenB strains, v2 FHbp accounted for 90.8%. 79.7% of clinical MenC strains were positive for v1 FHbp and 20.3% were positive for v2 FHbp. Among carrier-derived MenC strains, v2 FHbp predominated. The number of major serogroups of N. meningitidis analyzed by MLST was 822, and the encoded FHbp showed CC- or ST-specific characteristics. ConclusionfHbp genes were detected in almost all N. meningitidis strains in this study. Therefore, it is possible that a vaccine against MenB or meningococci irrespective of serogroups, which includes FHbp, could be developed. Meningococcal vaccine development for China is a complex issue and these findings warrant further attention with respect to vaccine development.

Detection of Neisseria meningitidis in asymptomatic carriers in a university hospital from Brazil

Asymptomatic meningococcus carriers in hospitals is a risk factor for acquiring meningococcal disease. Meningococcal carrier (MC) frequency was investigated in oropharyngeal swab samples collected from 200 staff members at a teaching hospital from Brazil. MC prevalence was 9% (95% CI 5–13%). Risk factors associated with MC were: mean age of 26.5 years, male gender, bar attendance frequency and number of persons/house. Of 18 isolated meningococcal strains, 14 were non-groupable (NG), 3 corrresponded to serogroup B and 1 to serogroup 29E. The frequency of serotypes and serosubtypes was heterogenous, with a slight predominance of serotypes 4 and 7 and serosubtypes P1.7 and P1.5. Most strains (n=13) were susceptible to the antimicrobials tested. The ctrA gene (PCR) was identified in 9 (64.3%) of the 14 NG strains, suggesting virulence in most of the NG isolated strains. Therefore, a constant surveillance of these asymptomatic carriers is required.

Meningococcal carriage among Georgia and Maryland high school students.

Meningococcal disease incidence in the United States is at an all-time low. In a previous study of Georgia high school students, meningococcal carriage prevalence was 7%. The purpose of this study was to measure the impact of a meningococcal conjugate vaccine on serogroup Y meningococcal carriage and to define the dynamics of carriage in high school students. This was a prospective cohort study at 8 high schools, 4 each in Maryland and Georgia, during a school year. Students at participating schools received quadrivalent meningococcal conjugate vaccine that uses diphtheria toxoid as the protein carrier (MCV4-DT). In each state, 2 high schools were randomly assigned for MCV4-DT receipt by students at the beginning of the study, and 2 were randomly assigned for MCV4-DT receipt at the end. Oropharyngeal swab cultures for meningococcal carriage were performed 3 times during the school year. Among 3311 students, the prevalence of meningococcal carriage was 3.21%-4.01%. Phenotypically nongroupable strains accounted for 88% of carriage isolates. There were only 5 observed acquisitions of serogroup Y strains during the study; therefore, the impact of MCV4-DT on meningococcal carriage could not be determined. Meningococcal carriage rates in US high school students were lower than expected, and the vast majority of strains did not express capsule. These findings may help explain the historically low incidence of meningococcal disease in the United States.

Carriage of Meningococci by University Students, United Kingdom

To the Editor: Neisseria meningitidis causes septicemia and meningitis (1). Meningococci usually persist on the nasopharyngeal mucosa of asymptomatic carriers (2). Because carriers are the only reservoir of meningococci, carriage in at-risk populations should be monitored. Meningococcal carriage rates have been assessed during 1997–8 for first-year students at the University of Nottingham (3) and in autumn during 1999–2001 for >48,000 sixth-form students (pre-university, age range 15–17 years) throughout the United Kingdom (4). Serogroup B and nongroupable strains predominated; serogroup Y strains were found in only 1%–2% of participants. From November 2008 through May 2009, to investigate persistence and spread of meningococcal strains in students living in dormitories, we conducted a longitudinal study in a cohort of 190 first-year students at the University of Nottingham. We found high rates of carriage and prevalence of serogroup Y strains (5). During September 2009 (first week of term) through March 2010, we conducted a large repeated cross-sectional study analyzing pharyngeal swabs from students in all school-year groups at Nottingham University. The objective of this study was to determine the significance of changes in overall meningococcal and serogroup Y-specific carriage rates among students. In September, first-year students were recruited on the main campus during registration and subsequently in dormitories and the main library. Undergraduates not in the first year were all recruited in the main library. This September sample of 823 first-year students represents 16.5% of the 5,000 undergraduate students registered each academic year on the main campus. Although not intentional, some overlap occurred when students were resampled during subsequent visits to the same dormitories and library, e.g., among the 557 first-year students from whom swab samples were collected in December, 74 (13%) had previously provided swab samples. Our study was approved by the Nottingham University Medical School Ethics Committee, and written informed consent was obtained from all participants. Pharyngeal swab samples were spread onto GC selective agar (Oxoid, Basingstoke, UK) and incubated at 37°C in air containing 5% CO2. After 48 hours, colonies suggestive of Neisseria spp. were examined for positive oxidase reaction; single colonies were confirmed as meningococci by amplification of meningococcal genes crgA plus ctrA and/or porA (6). PCR-based serogrouping was performed as described (6,7). Chi-square tests for significance were performed by using STATCALC (Epi Info version 6.04; Centers for Disease Control and Prevention, Atlanta, GA, USA). Among first-year students, carriage rates increased from 23.2% in late September to 55.7% by mid-December and remained at a similar level in March (Table). Among second- and third-year students, carriage rates were 34.2% and 30.5% in September, respectively, and remained at similar levels throughout the academic year. The increase in carriage among first-year students from September through December was mainly the result of a significant (23%) increase in carriage of serogroup Y strains (Table). In contrast, during the same period, carriage rates of serogroup Y strains did not change significantly among second- and third-year students (Table). Table Characteristics of meningococci carriage, University of Nottingham students, United Kingdom, 2009–10* Initial carriage rates were significantly higher for incoming (first-year) students in September 2009 than in 1997 (13.9% [3]; χ2 = 14, 1 df; p<0.0001); swabbing and culture protocols and sampling sites were identical in both studies, so the increases are real. Because 83% of students at Nottingham University come from all regions of the United Kingdom and 17% from other countries, the increased rates of carriage may reflect a nationwide change (8). Furthermore, testing within the first week of term meant that recovered strains were predominately brought into the university. Serogroup Y carriage rates for incoming students (2.9%) were significantly higher than rates detected by identical genotyping methods during 1999–2001 (1.7%–1.8% [4]; χ2 = 4.6%–6.4%, 1 df; p<0.05), suggesting that meningococcal carriage by young adults, particularly of serogroup Y strains, has increased across the United Kingdom. The major increase in serogroup Y strains among first-year students during 2009–10 probably resulted from spread of clones within dormitories, as observed in the 2008–9 study (5) and may be facilitated by characteristics of the organism, lack of immunity, or a combination of these factors. The high prevalence of serogroup Y strains in carriers may help explain the recent increased incidence of serogroup Y disease in the United Kingdom: from 20 to 62 laboratory-confirmed cases in England and Wales from 2003 through 2009 (9). In the United States during the late 1990s, a similar increase in serogroup Y carriage was linked to a concomitant increase in serogroup Y disease (10). In conclusion, in a representative UK student cohort we detected high rates of carriage and elevated prevalence of serogroup Y strains of meningococci. Any further significant increase in serogroup Y disease should lead to prompt reconsideration of the current vaccine policy in the United Kingdom.

Differential Expression and Transcriptional Analysis of the α-2,3-Sialyltransferase Gene in Pathogenic Neisseria spp

Alpha-2,3-sialyltransferase (Lst) is expressed on the outer membrane of Neisseria gonorrhoeae and Neisseria meningitidis and sialylates surface lipooligosaccharide (LOS), facilitating resistance to complement-mediated killing. The enzyme is constitutively expressed from a single gene (lst) and does not undergo antigenic or phase variation. We observed that Triton X-100 extracts of N. gonorrhoeae strain F62 contain about fivefold more sialyltransferase (Stase) activity than extracts of N. meningitidis strain MC58 [symbol: see text]3 a serogroup B acapsulate mutant. We confirmed and expanded upon this observation by showing that extracts of 16 random N. gonorrhoeae isolates contain various amounts of Stase activity, but, on average, 2.2-fold-more Stase activity than extracts of 16 N. meningitidis clinical isolates, representing several serogroups and nongroupable strains. Northern and real-time reverse transcription-PCR analysis of lst transcript levels in N. gonorrhoeae and N. meningitidis revealed that N. gonorrhoeae strains express more lst transcript than N. meningitidis strains. Although transcript levels correlate with average Stase activity observed in the two species, there was not a direct correlation between lst transcript levels and Stase activity among individual isolates of each species. Comparison of lst upstream (5'lst) regions of N. gonorrhoeae and N. meningitidis revealed striking sequence differences characteristic of the two pathogens. N. gonorrhoeae 5'lst regions possess 30-bp and 13-bp elements present as single elements or as tandem repeats that exist only as single elements in the 5'lst regions of N. meningitidis isolates. In addition, the 5'lst regions of N. meningitidis strains have 105-bp transposon-like Correia elements which are absent in N. gonorrhoeae. Chromosomal N. gonorrhoeae 5'lst::lacZ translational fusions expressed 4.75 +/- 0.09-fold (n = 4) higher beta-galactosidase (beta-gal) activity than N. meningitidis 5'lst::lacZ fusions in a host-independent manner, indicating differential expression is governed at least in part by sequence variations in the 5'lst regions. Reporter fusion assays and promoter-mapping analysis revealed that N. gonorrhoeae and N. meningitidis use different promoters with different strengths to transcribe lst. In N. gonorrhoeae, a strong sigma 70 promoter 80 bp upstream of the translational start site is used to transcribe lst, whereas this promoter is inactive in N. meningitidis. In N. meningitidis, a weak sigma 70 promoter at the 3' terminus of a 105-bp Correia repeat-enclosed element 99 bp upstream of the translational start site is used to transcribe lst. We conclude that differential Stase expression between N. gonorrhoeae and N. meningitidis is due at least in part to differential lst gene transcription.

A novel porA-based real-time PCR for detection of meningococcal carriage

Real-time PCR based on the capsule transfer gene (ctrA) is a significant aid in the diagnosis of meningococcal infection but fails to detect a high proportion (60 %) of non-groupable strains associated with nasopharyngeal carriage. This study aimed to design a novel real-time (TaqMan) PCR that would detect more strains of meningococci and be suitable for large-scale carriage studies. Primer and probe sequences were based on the meningococcal porA gene and designed specifically to exclude the highly related porA pseudogene in Neisseria gonorrhoeae. The specificity of the assay was confirmed by testing strains of N. gonorrhoeae known to contain the porA pseudogene together with commensal strains of Neisseria lactamica and Neisseria sicca. None of these was detected in the assay. Neisseria meningitidis strains representing a wide range of serogroups together with non-groupable strains isolated from the nasopharynx were tested by ctrA assay and the novel porA-based TaqMan PCR. All carriage strains were detected by the porA-based assay including four that gave weak or no reaction with the ctrA assay. Comparison of ctrA and porA assays on 71 throat swabs obtained from university students showed that the porA assay detected meningococcal DNA in all samples that were ctrA positive plus three that were ctrA negative but culture positive. This novel porA-based TaqMan assay provides a highly specific method for detecting meningococcal DNA that is more sensitive than the ctrA assay for detecting meningococcal carriage and is particularly suitable for carriage studies where non-groupable strains and other Neisseria are present.

Les arthrites septiques streptococciques de l’adulte : 55 cas et revue de la littérature

Objectives. – To evaluate the rate of occurrence and characteristics of streptococcal septic arthritis. Methods . – Retrospective single-center study of patients with bacteriologically documented septic arthritis admitted to a rheumatology department over a 20-year period. Results. – Of 303 cases of septic arthritis, 55 (18%) were due to streptococci and 166 (55%) to S. aureus (55%). As compared to patients with S. aureus arthritis, patients with streptococcal arthritis were more likely to be female (56% vs. 36%, P < 0.006) and older than 60 years of age (71% vs. 58%), less likely to have comorbidities (36% vs. 56%), rheumatoid arthritis (5% vs. 19%, P < 0.01), or diabetes (2% vs. 15%, P < 0.01), and more likely to have cancer (13% vs. 7%). Involved joints and proportions of patients with arthritis in multiple joints were similar in the two groups. Mortality was lower in the group with streptococcal infection (3.6% vs. 7.8%). The streptococci were distributed as follows: Group A ( n = 7), Group B ( n = 12), Group C ( n = 4), Group D ( n = 7), Group F ( n = 1), Group G ( n = 2), nongroupable ( n = 14), nontypable ( n = 1), and S. pneumoniae ( n = 7). Groups A and B and nongroupable strains mainly affected women; Group A selectively involved younger patients and Group B very elderly patients. Comorbidity, most notably cancer, was common in patients with S. pneumoniae or Group D streptococci. The portal of entry was often a skin lesion for Groups A and B and a medical procedure for Group D. Multiple joint involvement was common with Groups A and B and prosthetic joint infection with Groups B and C. Group A and S. pneumoniae were associated with severe systemic symptoms and extraarticular foci of infection, whereas a smoldering course was more common with Groups D and G and with nongroupable strains. Residual joint abnormalities were noted in half the patients, with no differences across groups. Conclusions . – The features of streptococcal septic arthritis vary according to the group of the causative organism and differ from those of S. aureus arthritis.

Streptococcal septic arthritis in adults. A study of 55 cases with a literature review

Objectives. – To evaluate the rate of occurrence and characteristics of streptococcal septic arthritis. Methods. – Retrospective single-center study of patients with bacteriologically documented septic arthritis admitted to a rheumatology department over a 20–year period. Results. – Of 303 cases of septic arthritis, 55 (18%) were due to streptococci and 166 (55%) to Staphylococcus aureus (55%). As compared to patients with S. aureus arthritis, patients with streptococcal arthritis was more likely to be in female (56% vs. 36%, P < 0.006) and older than 60 years of age (71% vs. 58%), less likely to have comorbidities (36% vs. 56%), rheumatoid arthritis (5% vs. 19%, P < 0.01), or diabetes (2% vs. 15%, P < 0.01), and more likely to have cancer (13% vs. 7%). Involved joints and proportions of patients with arthritis in multiple joints were similar in two groups. Mortality was lower in the group with streptococcal infection (3.6% vs. 7.8%). The streptococci were distributed as follows: group A ( n = 7), group B ( n = 12), group C ( n = 4), group D ( n = 7), group F ( n = 1), group G ( n = 2), nongroupable ( n = 14), nontypable ( n = 1), and Streptococcus pneumoniae ( n = 7). Groups A and B and nongroupable strains mainly affected women; group A selectively involved younger patients and group B very elderly patients. Comorbidity, most notably cancer, was common in patients with S. pneumoniae or group D streptococci. The portal of entry was often a skin lesion for groups A and B and a medical procedure for group D. Multiple joint involvement was common with groups A and B and prosthetic joint infection with groups B and C. Group A and S. pneumoniae were associated with severe systemic symptoms and extra articular foci of infection, whereas a smoldering course was more common with groups D and G and with nongroupable strains. Residual joint abnormalities were noted in half the patients, with no differences across groups. Conclusions. – The features of streptococcal septic arthritis vary according to the group of the causative organism and differ from those of S. aureus arthritis.

Neisseria meningitidis W-135 carriage during the Hajj season 2003.

During the 2003 Hajj pilgrimage to Mecca, 344 pilgrims of 29 different nationalities were screened by means of a throat swab to detect Neisseria meningitidis carriage. N. meningitidis was isolated from 11 subjects; 2 were serogroup W-135, 1 serogroup B, and 8 were non-groupable. The results indicate a very low colonization rate for N. meningitidis among the tested cohort, with a predominance of non-groupable strains. These results, combined with a review of the published data, warrant a re-evaluation of current recommendations by the Saudi Ministry of Health for the use of ciprofloxacin for Saudi pilgrims departing at the end of the Hajj season. However, vaccination with the meningococcal quadrivalent vaccine, for all pilgrims, should continue to be recommended. The possibility of new strains arising as a cause of future meningococcal outbreaks should be considered, and annual surveillance may give an early warning.

Dynamics of meningococcal long-term carriage among university students and their implications for mass vaccination.

In the 1997-98 academic year, we conducted a longitudinal study of meningococcal carriage and acquisition among first-year students at Nottingham University, Nottingham, United Kingdom. We examined the dynamics of long-term meningococcal carriage with detailed characterization of the isolates. Pharyngeal swabs were obtained from 2,453 first-year students at the start of the academic year (October), later on during the autumn term, and again in March. Swabs were immediately cultured on selective media, and meningococci were identified and serologically characterized. Nongroupable strains were genetically grouped using a PCR-based assay. Pulsed-field gel electrophoresis was used to determine the link between sequential isolates. Of the carriers initially identified in October, 44.1% (98 of 222) were still positive later on in the autumn (November or December); 57.1% of these remained persistent carriers at 6 months. Of the index carriers who lost carriage during the autumn, 16% were recolonized at 6 months. Of 344 index noncarriers followed up, 22.1% acquired carriage during the autumn term and another 13.7% acquired carriage by March. Overall, 43.9% (397 of 904) of the isolates were noncapsulated (serologically nongroupable); by PCR-based genogrouping, a quarter of these belonged to the capsular groups B and C. The ratio of capsulated to noncapsulated forms for group B and C strains was 2.9 and 0.95, respectively. Sequential isolates of persistent carriers revealed that individuals may carry the same or entirely different organisms at different times. We identified three strains that clearly switched their capsular expression on and off at different times in vivo. One student developed invasive meningococcal disease after carrying the same organism for over 7 weeks. The study revealed a high rate of turnover of meningococcal carriage among students. Noncapsulated organisms are capable of switching their capsular expression on and off (both ways) in the nasopharynx, and group C strains are more likely to be noncapsulated than group B strains. Carriage of a particular meningococcal strain does not necessarily protect against colonization or invasion by a homologous or heterologous strain.

Assessment of complement deficiency in patients with meningococcal disease in The Netherlands.

The frequency of complement deficiency in 176 of 7,732 patients with meningococcal disease in the Netherlands from 1959 through 1992 was assessed. Complement deficiency was found in six patients (3%): 3 (7%) of the patients with Neisseria meningitidis serogroup C disease, 1 (2%) of the patients with N. meningitidis serogroup A disease, and 2 (33%) of the patients with infections due to uncommon serogroups and nongroupable strains of N. meningitidis. Of 91 additional patients with meningococcal infections due to uncommon serogroups, 33% also had complement deficiency. Thirty-four of the 36 complement-deficient patients with meningococcal disease who were from 33 families were 5 years of age or older. Twenty-six additional complement-deficient relatives were found. Screening individuals with meningococcal disease due to uncommon serogroups who were 5 years of age or older identified 30 of the 33 complement-deficient families. Only 27% of the complement-deficient relatives had had meningococcal disease. This risk was lower for relatives with properdin deficiency (18%) than for those deficient in the late component of complement (38%). Therefore, pedigree studies are warranted for identifying those complement-deficient persons who require vaccination for meningococcal disease.

Evaluation of the chick embryo for the determination of relative virulence of Neisseria meningitidis

The chick embryo model was evaluated as a method to compare virulence between selected strains of Neisseria meningitidis. Inoculation of 13-day-chick embryos via the egg yolk distinguished strains having an LD 50 of 10 3 colony forming units (CFU) or greater (low virulence) from those having an LD 50 of approximately 10 1 or less (high virulence). A strain of serogroup B and a spontaneous nonpiliated strain of group C were found to be of relatively high virulence while a strain of N. lactamica, a serogroup A carrier strain, and certain nongroupable strains were found to be of low virulence. Strains having an LD 50 of 10 2 were not differentiated from either of these. Alternatively, inoculation of the chorioallantoic membrane (CAM) of 9-day-old chick embryos statistically differentiated most strains of N. meningitidis although inoculation via this route was less sensitive.

Evaluation of the chick embryo for the determination of relative virulence ofNeisseria meningitidis

The chick embryo model was evaluated as a method to compare virulence between selected strains of Neisseria meningitidis. Inoculation of 13-day-chick embryos via the egg yolk distinguished strains having an LD50 of 10(3) colony forming units (CFU) or greater (low virulence) from those having an LD50 of approximately 10(1) or less (high virulence). A strain of serogroup B and a spontaneous nonpiliated strain of group C were found to be of relatively high virulence while a strain of N. lactamica, a serogroup A carrier strain, and certain nongroupable strains were found to be of low virulence. Strains having an LD50 of 10(2) were not differentiated from either of these. Alternatively, inoculation of the chorioallantoic membrane (CAM) of 9-day-old chick embryos statistically differentiated most strains of N. meningitidis although inoculation via this route was less sensitive.

Sero/subtyping of Neisseria meningitidis isolated from patients in Spain.

To know the types of meningococcal strains in Spain, we serotyped and subtyped 743 Neisseria meningitidis isolates recovered between 1990 and 1992 from patients. A great number of serogroup B, serogroup C and non-groupable meningococci reacted with the serotyping reagents while many serogroup C and non-groupable isolates did not react with the serosubtyping reagents (78.2% and 54.8% respectively); only 8.9% of serogroup B meningococci were non-subtypeable (NST). Distribution of serotypes was similar in serogroup C and in non-groupable strains. Isolates showed great variability in antigenic phenotypes (71 in serogroup B, 20 in serogroup C and 25 in non-groupable meningococci). The most frequent antigenic combinations were 4:P1.15 (39.8%) in serogroup B, 2b:NST (55.8%) in serogroup C and 2b:NST (35.6%) in non-groupable meningococci.

Complement deficiency predisposes for meningitis due to nongroupable meningococci and Neisseria-related bacteria.

Nongroupable meningococci or bacteria related to the genus Neisseria rarely cause meningitis. Complement deficiency has been identified as a major predisposing factor for meningococcal disease. To assess whether patients with meningitis due to such strains have a complement deficiency, we studied 12 persons. Six patients had meningitis due to nongroupable strains of meningococci, and six patients had meningitis due to Moraxella species or Acinetobacter species. Inherited complement component C7 or C8 deficiency was found in two persons who had had meningitis due to nongroupable meningococci, and one C8-deficient person had had meningitis caused by Moraxella osloensis. Hypocomplementemia resulting from CSF drain-associated shunt nephritis was found in one person with meningitis due to Moraxella nonliquefaciens and in one person with meningitis due to Acinetobacter lwoffi. This rather high frequency of inherited or acquired complement deficiencies among patients with meningitis due to nongroupable meningococci, Moraxella species, and Acinetobacter species justifies the recommendation that such patients must be studied for complement deficiency.

Carriage of Neisseria meningitidis and Neisseria lactamica in a school population during an epidemic period in Spain.

A study was made of the incidence of Neisseria meningitidis and N. lactamica in a school population; 2470 children aged between 5 and 7 years were studied from four schools in Alcala de Henares (Madrid). Nasopharyngeal swabs were taken in June, November and March, between 1979 and 1983. In all the surveys except one, the proportion of carriers of N. lactamica was higher than that of N. meningitidis, reaching a ratio of about 2:1 in the complete study. The predominant serogroup of meningococcus found was B (41%), with nongroupable strains reaching 43%. A study of serotypes within group B showed a predominance of nontypable strains (48.5%), while those strains considered to be most virulent (types 2 and 1, 8, 15) reached 40%. Eighteen per cent of N. lactamica strains were observed to agglutinate with antimeningococcal sera whilst the remainder of the strains were rough. When these strains were studied with the antiserum-agar technique, using antimeningococcal sera, a high percentage of strains cross-reacted with the meningococci. The susceptibility of strains to sulphadiazine, penicillin, ampicillin, chloramphenicol, rifampicin and spiramycin was determined. Finally an analysis was made of the effect that an elevated colonization rate of N. lactamica might have on colonization by meningococci. The necessity of using fine epidemiological markers in tracing virulent strains in a population at risk is stressed. Selective prophylactic measures are also necessary.